. 2016 Jun 7;15:98. doi: 10.1186/s12934-016-0499-2

Table 2.

Methoxine incorporation in the Mut^S and O-acetyl homoserine sulfhydrylase knock-out strains

Strain	Genotype	Source	Nanobody B
Strain	Genotype	Source	Titers (g l⁻¹)	% Δ−16 Da variant
CBS7435Mut^S	aox1	[7]	2.4 ± 0.7	0.90 ± 0.17
CBS7435Mut^SΔOAHS	aox1 met25	This study	2.1 ± 0.2	0

Nanobody B was produced in the CBS7435Mut^S and in the CBS7435Mut^S strain with an inactivation of O-acetyl homoserine sulfhydrylase gene (CBS7435Mut^SΔOAHS) in shake flasks using complex media. Titers of Nanobody B were analyzed via a proteinA cleanup step followed by OD₂₈₀ measurement. Relative abundance of the methoxine variant was analysed via RP-HPLC followed by total mass measurement by ESI-Q-TOF-MS. The methoxine variant of Nanobody B was only produced in the Mut^S strain and absent when O-acetyl homoserine sulfhydrylase was inactivated. Experiments were performed in triplicate